SALIVARY MARKERS OF SYSTEMIC DISEASES<br />BY<br />Y.PRAGZNA(10T21S0113)<br />UNDER THE GUIDANCE OF<br />Mrs. M. SANDHYA M...
CONTENTS:<br /><ul><li>Biomarkers
Saliva physiology
Advantages & disadvantages
Saliva collection, processing & storage
Diagnostic applications
Salivary proteome & transcriptome
New technologies of measurement</li></li></ul><li>BIOMARKERS:<br />Biomarkers are defined as cellular, biochemical and mol...
SALIVARY MARKERS<br />Non invasive diagnosis of disease and monitoring of general health<br />
SALIVA PHYSIOLOGY:<br /><ul><li>Saliva is the mixed product of 3 major salivary glands.</li></ul>                         ...
Several physiological & pathological  conditions  modify  saliva  production quantitatively.</li></li></ul><li>Functions o...
Anti-microbial
Mucosal integrity
Lavage/cleansing
Buffering
Remineralization</li></ul>FOOD & SPEECH RELATED<br /><ul><li>Food preparation
Digestion
Taste
Speech
Prevention of caries</li></li></ul><li>SALIVA  AS  A DIAGNOSTIC FLUID<br />ADVANTAGES<br /><ul><li>It is safe, non-invasiv...
No needles and No cringing.
Home testing.
It has an average protein concentration of 1.5-2 mg/ml.
Cost of shipping & Storage tend to be lower.</li></ul>Informative analytes are present in lower amounts in saliva than in ...
Whole saliva collection<br />Ductal secretions collection<br />Parotid secretions collection<br />Sub mandibular & sub lin...
PROCESSING & STORAGE:   <br />All procedures must be performed on ice:<br />(1)Divide the saliva samples into multiple 330...
RNA analysis: Add 1.65µL of SUPERase inhibitor.</li></ul>(2)Invert gently to mix.<br />(3)Store all fractions at -80ºc.<br />
Salivary/Oral Fluid Biomarkers Possibilities of Use <br />DNA                     Standard genotyping<br />               ...
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Salivary markers of systemic diseases ppt

  1. 1. SALIVARY MARKERS OF SYSTEMIC DISEASES<br />BY<br />Y.PRAGZNA(10T21S0113)<br />UNDER THE GUIDANCE OF<br />Mrs. M. SANDHYA M.PHARM<br />ASSOCIATE PROFESSOR<br />
  2. 2. CONTENTS:<br /><ul><li>Biomarkers
  3. 3. Saliva physiology
  4. 4. Advantages & disadvantages
  5. 5. Saliva collection, processing & storage
  6. 6. Diagnostic applications
  7. 7. Salivary proteome & transcriptome
  8. 8. New technologies of measurement</li></li></ul><li>BIOMARKERS:<br />Biomarkers are defined as cellular, biochemical and molecular characteristics by which normal/abnormal processes can be recognized and/or monitored.<br />
  9. 9. SALIVARY MARKERS<br />Non invasive diagnosis of disease and monitoring of general health<br />
  10. 10. SALIVA PHYSIOLOGY:<br /><ul><li>Saliva is the mixed product of 3 major salivary glands.</li></ul> Parotid<br /> Sub mandibular<br /> Sub lingual<br /><ul><li>It is slightly acidic(pH 6-7).
  11. 11. Several physiological & pathological conditions modify saliva production quantitatively.</li></li></ul><li>Functions of saliva<br />PROTECTIVE<br /><ul><li>Lubrication
  12. 12. Anti-microbial
  13. 13. Mucosal integrity
  14. 14. Lavage/cleansing
  15. 15. Buffering
  16. 16. Remineralization</li></ul>FOOD & SPEECH RELATED<br /><ul><li>Food preparation
  17. 17. Digestion
  18. 18. Taste
  19. 19. Speech
  20. 20. Prevention of caries</li></li></ul><li>SALIVA AS A DIAGNOSTIC FLUID<br />ADVANTAGES<br /><ul><li>It is safe, non-invasive and simple.
  21. 21. No needles and No cringing.
  22. 22. Home testing.
  23. 23. It has an average protein concentration of 1.5-2 mg/ml.
  24. 24. Cost of shipping & Storage tend to be lower.</li></ul>Informative analytes are present in lower amounts in saliva than in serum.<br />DISADVANTAGES<br />
  25. 25. Whole saliva collection<br />Ductal secretions collection<br />Parotid secretions collection<br />Sub mandibular & sub lingual secretions collection<br />Saliva collection <br />
  26. 26. PROCESSING & STORAGE: <br />All procedures must be performed on ice:<br />(1)Divide the saliva samples into multiple 330μL samples placed in cryotubes at -80ºc.<br />Further processed for <br /><ul><li>Protein analysis: Add protease inhibitors like aprotinin(0.33µL), 3.3µL PMSF.
  27. 27. RNA analysis: Add 1.65µL of SUPERase inhibitor.</li></ul>(2)Invert gently to mix.<br />(3)Store all fractions at -80ºc.<br />
  28. 28. Salivary/Oral Fluid Biomarkers Possibilities of Use <br />DNA Standard genotyping<br /> Genetic information of the hosting human body <br /> Bacterial infection (oral microbes in the mouth) Diagnosing carcinomas of head & neck<br /> Forensics <br />mRNA Information on transcription rates<br />RNA Viral/bacterial identification<br />Proteins Genetic information<br /> Translational regulation <br /> Diagnosing periodontitis<br /> Detecting dental cavities<br />Immunoglobulin Diagnosing viruses <br />Metabolites Diagnosing periodontitis <br />
  29. 29. DIAGNOSTIC APPLICATIONS OF SALIVA:<br /><ul><li>Drugs, hormones:
  30. 30. Intra cellular pathways</li></ul> Active & Passive diffusion<br /><ul><li>Extra cellular pathways</li></ul> Ultra filtration<br /><ul><li>Serum components:
  31. 31. Capillary barrier
  32. 32. Interstitial spaces
  33. 33. Cervicular fluid
  34. 34. Membrane of acinar </li></ul> & ductal cells<br />
  35. 35. <ul><li>SJOGRENS SYNDROME:</li></ul>There are raised concentrations of NaCl, IgA, IgG, lactoferrin, inflammatory mediators such as PGE2, TXB2, Interleukin-6 etc.<br /><ul><li>CYSTIC FIBROSIS:</li></ul>Saliva of CF patients contain increased calcium & phosphate levels, urea & uric acid and abnormally elevated levels of PGE2.<br /><ul><li>DIABETES MELLITUS:</li></ul>It effects flow rate & composition of saliva. Salivary alterations includes higher levels of IgA, Salivary Peroxidase, glucose content, K, Salivary total protein & amylase.<br /><ul><li>ORAL MUCOSAL DISEASES:</li></ul>Nitric oxide levels were increased.<br />
  36. 36. <ul><li>ONCOLOGY:</li></ul>P53 antibody can be detected in oral squamous cell carcinoma, increased levels of salivary kallikrein levels in oral malignant tumors, CA 15-3 in breast cancer. CVS:<br />Determination of salivary amylase activity before 6hrs after cv surgery.<br /> KIDNEY DYSFUNCTION: <br />Hypo salivation , changes in taste, ammonium smelling breath, oral mucosal pain.<br />EPILEPSY:<br />Gingival hypertrophy, IgA deficiency<br />
  37. 37. VIRAL DISEASES:Acute hepatitis A & B were diagnosed based on the <br />presence of IgM antibodies in saliva.HIV:Salivary IgA levels.BACTERIAL INFECTIONS:By monitoring the presence of antibodies to microorganisms found in saliva & oral cavity<br />
  38. 38. <ul><li>AGAIN & AGAIN ABOUT TOBACCO:
  39. 39. Smoking causes decrease in salivary important antioxidant levels.
  40. 40. Salivary nicotine levels are considered to be the most reliable marker</li></li></ul><li>Drug monitoring<br />ANTI-ASTHMATICS:Theophylline<br />ANTI EPILEPTICS: Carbamazapine, Diazepam, Ethosuximide, Lamotrigine, Topiramate, Phenytoin, Primidone, Valproic acid<br />ANTI ARRYTHMIC/ANTI HYPERTENSIVES: Digoxin, Metoprolol, and Procainamide.<br />ANTI MICROBIALS:Clarithromycin, Gentamycin, Isoniazid, Ofloxacin, Sulfanilamide<br />ANTIPYRETICS: Acetaminophen, Antipyrine, Paracetamol<br />ANTI NEOPLASTICS:Cisplatin, Methotrexate, Texol, Topotecan<br />ANTI VIRAL: Quinine<br />LITHIUM PSYCHOTROPIC:Lithium, Amitriptyline<br />MISCELLANEOUS: Nicotine, Ethanol, Caffeine, Tolbutamide, Quinine<br />ILLICIT DRUGS: Amphetamine, Barbiturates, Benzodiazepines, Opiods, Cocaine, Phencyclidine<br />
  41. 41. HORMONE MONITORING:<br />Steroids like cortisol, aldosterone, testosterone, insulin<br />SIALOCHEMISTRY ANALYSIS:<br />Early monitoring of an exposure to lead, cadmium, mercury because of higher salivary levels of these elements<br />
  42. 42. CLASSIFICATION OF SALIVARY BIO MARKERS:<br /><ul><li>PROTEOMIC
  43. 43. GENOMIC
  44. 44. MICROBIAL
  45. 45. OTHERS</li></li></ul><li>Diagnostic molecular targets<br />SALIVARY PROTEOME:<br />Use of ‘Divide & conquer’ strategy<br /><ul><li>Initial fractionation of proteins with separation techniques
  46. 46. Digestion with proteolytic enzyme, trypsin
  47. 47. Analysis by 1-D, 2-D LC-MS
  48. 48. Finally data is processed & submitted for database searching</li></li></ul><li>SALIVARY PROTEOMIC ANALYSIS<br />
  49. 49. SALIVARY TRANSCRIPTOME:<br /><ul><li>It includes mRNA molecules that cells use to convey the instructions carried by DNA for subsequent protein production.
  50. 50. At present, the main strategy for identification of salivary transcriptomic biomarkers is through microarray technology.</li></li></ul><li>SALIVARY TRANSCRIPTOME:<br />DNA MICROARRAY TECHNOLOGY<br />
  51. 51. NEW TECHNOLOGIES FOR MEASURING SALIVARY BIOMARKERS:<br /><ul><li>POC DEVICE</li></ul>In this POC device, saliva(100-300μL) is placed into the salivary collection module, and then delivered into the Nano-biochip.<br /><ul><li>MICROBEAD ARRAY</li></ul>Micro pits within a silicon wafer are populated with a variety of chemically sensitized bead micro reactors.<br />
  52. 52. MICRO BEAD ARRAY TECHNIQUE<br />NANO-BIOCHIP<br />
  53. 53. CONCLUSION:<br /> The components of saliva act as a “Mirror of the body’s health”,<br /> widespread use and growing acceptability of saliva as a<br /> diagnostic tool is helping individuals, researchers, health care<br /> professionals and community health programs to better detect<br /> and monitor diseases and to improve the general health of the<br /> public<br />
  54. 54. REFERENCES:<br /><ul><li>Mandel ID. A contemporary view of salivary research. Crit Rev Oral Biol Med 1993; 4:599.
  55. 55. Raymond G, Schipper a, Erika Silletti a, Monique H, Vingerhoeds. Saliva as research material: biochemical, physiochemical, and practical aspects. Archives of Oral Biology 2007 ;( 52):1114-35.
  56. 56. Richard P, Jiri S, Jana V, Edgar F, Petr M, Jindrich P et al. Saliva as a diagnostic medium. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2009; 153(2):103-10.
  57. 57. Eliaz Kaufman .The diagnostic applications of saliva. A review. Crit Rev Oral Biol Med 2002; 13(2):197-212.
  58. 58. www.med.kufauniv.com/ar/journals2010/res%20(1).pdf
  59. 59. www.jofs.in/files/Artical-11.pdf
  60. 60. www.linkgroup.hu/docs/pcs2/218-08CurrGen.pdf
  61. 61. www.wikipedia.com</li></ul> <br />
  62. 62. THANK YOU<br />
  63. 63. QUERIES???<br />
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